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<p>A visual and dual-targets detection platform for <i>Staphylococcus aureus</i> and <i>Vibrio parahaemolyticus</i> was established based on recombinant polymerase amplification (RPA), CRISPR/Cas12a and lateral flow test strip. The test strip was constructed using multicolored microspheres labeling antibody as tracer, and the results of detection of two kinds of bacteria could be displayed by different colored test lines on the test strip through the antigen-antibody specific recognition function, which realized results visualization and avoid cross-reaction. Due to the target-specific amplification of RPA and the sequence-specific recognition of Cas12a, the detection strategy could still show good sensitivity, specificity and accuracy in complex matrices. The visual limit of detection was as low as 50 CFU/mL, and the entire detection process could be completed within 42&#xa0;min. In conclusion, a rapid and efficient detection platform for <i>Staphylococcus aureus</i> and <i>Vibrio parahaemolyticus</i> was developed, and this method is expected to provide a solution for the on-site detection of other foodborne pathogens in market and environmental food by simply changing the labeled antibody.</p> Graphical abstract <p></p>

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Red and blue microspheres based immunoassay combined with RPA and Cas12a for dual-detection of nucleic acid from foodborne pathogens

  • 期刊论文

  • 发表日期：

    2026年1月28日

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  • Lemei Zhu,
  • Jinbin Wang,
  • Xi Li,
  • Hui Zhen,
  • Ziyi Gu,
  • Laifa Wang,
  • Pingping Bing,
  • Haijuan Zeng

摘要

A visual and dual-targets detection platform for Staphylococcus aureus and Vibrio parahaemolyticus was established based on recombinant polymerase amplification (RPA), CRISPR/Cas12a and lateral flow test strip. The test strip was constructed using multicolored microspheres labeling antibody as tracer, and the results of detection of two kinds of bacteria could be displayed by different colored test lines on the test strip through the antigen-antibody specific recognition function, which realized results visualization and avoid cross-reaction. Due to the target-specific amplification of RPA and the sequence-specific recognition of Cas12a, the detection strategy could still show good sensitivity, specificity and accuracy in complex matrices. The visual limit of detection was as low as 50 CFU/mL, and the entire detection process could be completed within 42 min. In conclusion, a rapid and efficient detection platform for Staphylococcus aureus and Vibrio parahaemolyticus was developed, and this method is expected to provide a solution for the on-site detection of other foodborne pathogens in market and environmental food by simply changing the labeled antibody.

Graphical abstract